Dihydrochalcones: evaluation as novel radical scavenging antioxidants

Dihydrochalcones are a family of bicyclic flavonoids, defined by the presence of two benzene rings joined by a saturated three carbon bridge. In the present study, we systematically examined the antioxidant activities of dihydrochalcones against the stable free radical (1,1-diphenyl-2-picrylhydrazyl) and lipid peroxidation in the erythrocyte membrane. All dihydrochalcones exhibited higher antioxidant activities than the corresponding flavanones. The (1)H NMR analysis indicated that the active dihydrochalcone has a time-averaged conformation in which the aromatic A ring is orthogonal to the carbonyl group, while the inactive dihydrochalcone such as 2'-O-methyl-phloretin has a strongly hydrogen-bonded phenolic hydroxyl group, suggestive of a coplanar conformation. A hydroxyl group at the 2'-position of the dihydrochalcone A ring, newly formed by reduction of the flavanone C ring, is an essential pharmacophore for its radical scavenging potential.     

Application of the porapak q column extraction method for tomato flavor volatile analysis

The Porapak Q column method (PQM) was compared to the method of simultaneous distillation extraction (SDE) under reduced pressure for extraction of the volatile compounds produced by tomato cv. Momotaro. The PQM was found to be effective at trapping and isolating many low and high boiling point volatile compounds and at producing the very desirable natural ripe tomato flavor of extracts. The SDE method was less effective in isolating the higher boiling point volatile compounds and caused deterioration of volatile compounds due to the heating process that takes place during extraction, resulting in an unpleasant boiled green tomato flavor of extracts. The advantages of using the PQM are its simplicity and its high efficiency in isolating many volatile compounds from nonvolatile materials at room temperature. A total of 367 volatile compounds were isolated by the PQM. Of these, hexanal, (Z)-3-hexenal, (E)-2-hexenal, 2- and 3-methylbutanol, and 2-phenylethanol were relatively more abundant than other compounds and (Z)-3-hexenal showed the highest relative amount.     

Studies on replicating patterns of bovine sex chromosomes using 5–bromo-deoxyuridine (BrdU): I. The frequency of cell division and the length of cell cycle

Prior to the studies on the replicating patterns in bovine sex chromosomes, the frequency of cell division by the autoradiographic method of labeling with ³H-thymidine and the length of the cell cycle by the method of administration of 5–bro-modeoxyuridine (BrdU were examined in the cultured lymphocytes. The number of the cells labeled with ³H-thymidine increased punctually, and they almost attained their maximum number at 72 hours' labeling. In the cells to which BrdU had been added for over 36 hours, many cells passed through the cell cycle 3 times, and in the cells treated from 12 to 24 hours, almost all of the cells passed through the cell cycle two times. In the cells treated within 10 hours, almost none of the cells attained a full cell cycle. These results suggested that the length of the cell cycle in bovine cultured cells is about 12 hours .     

Transformed phenotypes in long-term cultures persistently infected with bovine leukemia virus.

Two cell lines, 3178 and FLK, were established respectively from calf form bovine lymphosarcoma and from fetal lamb kidney cells inoculated in vitro with bovine leukemia virus. These two cell lines persistently infected with bovine leukemia virus were maintained for more than 150 passages over three years. They exhibited the characteristics of transformed cell lines in vitro: 1) anchorage independence, 2) increased saturation density and decreased population doubling time, 3) increased uptake of 2-deoxy-D-glucose and 4) tumorigenicity in athymic nude mice.     

Sequence variants at the myostatin gene locus influence the body composition of Thoroughbred horses

Myostatin is a member of the transforming growth factor-β family with a key role in inhibition of muscle growth by negative regulation of both myoblast proliferation and differentiation. Recently, a genomic region on ECA18, which includes the MSTN gene, was identified as a candidate region influencing racing performance in Thoroughbreds. In this study, four SNPs on ECA18, g.65809482T>C, g.65868604G>T, g.66493737C>T, and g.66539967A>G, were genotyped in 91 Thoroughbred horses-in-training to evaluate the association between genotype and body composition traits, including body weight, withers height, chest circumference, cannon circumference, and body weight/withers height. Of these, statistically differences in body weight and body weight/withers height were associated with specific genotypes in males. Specifically, body weight/withers height showed statistically significant differences depending on genotype at g.658604G>T, g.66493737C>T, and g.66539967A>G (P<0.01) in males during the training period. Animals with a genotype associated with suitability for short-distance racing, C/C at g.66493737C>T, had the highest value (3.17 ± 0.05 kg·cm(-1)) for body weight/withers height in March, while those with a genotype associated with suitability for long-distance racing, T/T, had the lowest (2.99 ± 0.03 kg·cm(-1)). In females, the trends in the association of body weight/withers height with genotypes were similar to those observed in males. As the SNPs are not believed to be linked to coding variants in MSTN, these results suggest that regulation of MSTN gene expression influences skeletal muscle mass and hence racing performance, particularly optimum race distance, in Thoroughbred horses.     

Production of calcium maintenance factor Stanniocalcin-1 (STC1) by the equine endometrium during the early pregnant period

A factor responsible for progression to pregnancy establishment in the mare has not been definitively characterized. To identify factors possibly involved in the establishment of equine pregnancy, the endometrium was collected from day 13 (day 0=day of ovulation) cyclic and day 13, 19 and 25 pregnant animals. From initial subtractive hybridization studies, a calcium regulating factor, Stanniocalcin-1 (STC1) mRNA, was found as a candidate molecule expressed uniquely in the pregnant endometrium. Endometrial expression of STC1 mRNA was noted on day 19 and was markedly increased in the day 25 gravid endometrium. STC1 protein was found in the extracts of day 25 gravid endometrium and immunochemically localized in the uterine glands. In addition, STC1 protein was detected in uterine flushing media collected from day 25 pregnant mares. High concentrations of estradiol-17 β (E(2)) were detected in day 25 conceptuses. E(2) levels were much higher in the gravid endometrium than in other regions, whereas progesterone levels did not differ among the samples from different endometrial regions. Expression of STC1 mRNA, however, was not significantly upregulated in cultured endometrial explants treated with various concentrations of E(2) (0.01-100 ng/ml) with or without 10 ng/ml progesterone. These results indicate that an increase in STC1 expression appears to coincide with capsule disappearance in the conceptus, and suggest that STC1 from the uterine glands likely plays a role in conceptus development during the pregnancy establishment period in the mare.     

Antioxidative catechol lignans converted from sesamin and sesaminol triglucoside by culturing with Aspergillus

Sesamin and sesaminol triglucoside in sesame seeds are major lignans that display an abundance of biological activities. Although their antioxidative activity in vitro is weak, they have been reported to suppress oxidative stress in vivo. We investigated the production of new antioxidative lignans from sesame lignans by culturing with the genus Aspergillus to enhance the function of food materials. Media containing sesamin or sesaminol triglucoside increased antioxidative activity for DPPH radical scavenging by culturing with Aspergillus usamii mut. shirousamii RIB2503. The antioxidative lignans in sesamin medium were identified as sesamin 2,6-dicatechol and episesamin 2,6-dicatechol. Those in sesaminol triglucoside medium were identified as sesaminol 6-catechol and episesaminol 6-catechol, which are novel antioxidative lignans. It is suggested that they may exhibit higher antioxidative activity than sesamin and sesaminol triglucoside because they have the catechol functional moiety.     

Population of Follicles and Luteal Structures during the Oestrous Cycle of Mares Detected by Three-Dimensional Internal Structure Microscopy

The structure of the equine ovary is different from that of other mammals in its extremely large size, the presence of ovarian fossa and the inverted location of its cortex and medulla. A three-dimensional internal structure microscopy (3D-ISM), which consists of a computer-controlled slicer, a CCD camera, a laser disc recorder and a PC, is very useful for the observation of the internal structures in equine ovaries. In addition, the three-dimensional images of follicles and corpus luteum (CL) reconstructed by the segmentation technique can clarify the spatial arrangement in the equine ovary. In this study, to understand the changes in the ovarian internal structures of the mare during the oestrous cycle, the size and numbers of follicles and luteal structures were analysed by 3D-ISM in addition to the concentrations of progesterone (P₄) and oestradiol-17β. As a result, many small follicles (<10 mm in diameter) were detected. It was recognized that the luteal structures were distinguished into three types, such as the corpus haemorragicum (CH), which is formed by blood elements at the cavity after ovulation, CL and corpus albican (CA). There were some CHs and CL in the group, which had the concentration of P₄ > 1 ng/ml. CHs were also observed in the group, which had low level of P₄ (P₄ < 1 ng/ml). CAs were found regardless of the P₄ level. In conclusion, 3D-ISM enabled the internal observation of the ovarian structures in detail, and estimation of the stage of the ovarian cycle with complementary physiological information. The findings by 3D-ISM provide basic information for clinical applications.